CN-121736110-B - PR-resistant recombinant rabbit monoclonal antibody and application thereof

Abstract

The invention belongs to the technical field of immunochemistry, and particularly relates to an anti-PR recombinant rabbit monoclonal antibody and application thereof, wherein the anti-PR recombinant rabbit monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 4, and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 5. The invention also relates to a nucleotide sequence, a recombinant plasmid or an expression vector for encoding the PR-resistant recombinant rabbit monoclonal antibody, a preparation method, application of the PR-resistant recombinant rabbit monoclonal antibody in a PR protein detection method or device and the like. The PR recombinant rabbit monoclonal antibody has the characteristics of good specificity, strong positive signals and the like, is easier to score in IHC staining, and is more accurate for detecting and distinguishing cancers.

Inventors

• Xiao Muzhang
• Kang Fanhua
• LI JIANBO
• ZHANG DONGXU

Assignees

• 苏州百道医疗科技有限公司
• 中南大学湘雅医院
• 湘雅常德医院

Dates

Publication Date

20260508

Application Date

20260228

Claims (10)

1. 1. The anti-PR protein recombinant rabbit monoclonal antibody is characterized by comprising a heavy chain variable region and a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 4, and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 5.
2. 2. A coding gene for the recombinant rabbit monoclonal antibody against PR protein of claim 1.
3. 3. The coding gene according to claim 2, comprising a DNA sequence shown as SEQ ID NO. 2 for encoding a heavy chain variable region of the recombinant rabbit monoclonal antibody against PR protein and a DNA sequence shown as SEQ ID NO. 3 for encoding a light chain variable region of the recombinant rabbit monoclonal antibody against PR protein.
4. 4. A nucleic acid molecule comprising the coding gene of claim 2 or 3.
5. 5. An expression vector comprising the nucleic acid molecule of claim 4.
6. 6. A host cell transformed or transfected with the expression vector of claim 5.
7. 7. A preparation method of an anti-PR protein recombinant rabbit monoclonal antibody is characterized in that the expression vector of claim 5 is adopted to transform or transfect host cells, the transformed or transfected cells are cultured, cell supernatant is collected and purified, and the anti-PR protein recombinant rabbit monoclonal antibody is obtained.
8. 8. Use of the recombinant rabbit monoclonal antibody against PR protein according to claim 1, the encoding gene according to claim 2 or 3, the nucleic acid molecule according to claim 4, the expression vector according to claim 5, the host cell according to claim 6 for the preparation of a PR protein detection device.
9. 9. A PR protein detection kit comprising the recombinant rabbit monoclonal antibody against PR protein of claim 1.
10. 10. The PR protein detection kit according to claim 9, further comprising horseradish peroxidase labeled secondary antibody, ethylenediamine tetraacetic acid repair liquid, catalase blocking liquid, 3 '-diaminobenzidine concentrate, 3' -diaminobenzidine buffer, hematoxylin and bluing liquid.

Description

PR-resistant recombinant rabbit monoclonal antibody and application thereof Technical Field The invention belongs to the technical field of immunochemistry, and particularly relates to an anti-PR recombinant rabbit monoclonal antibody and application thereof, in particular to application in immunohistochemical detection. Background PR protein, namely the progestogen receptor (Progesterone Receptor, PR for short), is a core transcription factor mediating the biological effects of progestogen (Progesterone, P4) and has two subtypes, namely PR-A subtype with molecular weight of about 94kDa and PR-B subtype with molecular weight of about 110kDa. PR is a dual-function protein that can act as a transcription factor in a ligand-dependent manner to regulate transcription directly with nuclear DNA, and can act as a non-transcription factor to regulate an extranuclear signaling pathway. PR protein is combined with progestogen to regulate downstream gene expression, and is widely involved in female reproductive system development (such as uterus and mammary gland), pregnancy maintenance, embryo implantation and other physiological processes, and plays a key role in mediating and regulating functions of ovary, uterus and mammary gland and reproductive activities. The research shows that PR dysfunction is closely related to the occurrence and development of various reproductive system diseases and hormone-dependent tumors, has critical pathological significance in breast cancer and endometrial diseases, and is an important molecular marker for disease diagnosis, typing and treatment strategy formulation. At present, an Immunohistochemical (IHC) method is often used clinically to carry out tissue in-situ detection on PR proteins, and the expression condition of the PR proteins in cells is observed through a microscope, so that whether the PR proteins have abnormal expression or not is estimated. However, the anti-PR antibodies used in the prior art have obvious disadvantages in terms of sensitivity and specificity, and particularly have poor performance in distinguishing PR-A from PR-B subtypes, identifying low-expression samples, reducing nonspecific staining, and the like. The IHC staining result has the problems of strong subjectivity, poor repeatability and the like in pathological evaluation, and influences the accuracy of disease typing and treatment decision based on PR state. Therefore, the development of the anti-PR antibody which has high sensitivity and strong specificity and is suitable for pathological detection scenes such as IHC and the like has important practical significance for realizing accurate identification of PR proteins, improving the reliability of related disease diagnosis and promoting the establishment of individual treatment strategies. Disclosure of Invention First, the technical problem to be solved In view of the above-mentioned shortcomings and disadvantages of the prior art, the present invention provides an anti-PR recombinant rabbit monoclonal antibody which has a wide application range and can accurately recognize PR expression, and its application. The invention also relates to a nucleotide sequence, a recombinant plasmid or an expression vector for encoding the PR-resistant recombinant rabbit monoclonal antibody, a preparation method, application of the PR-resistant recombinant rabbit monoclonal antibody in a PR protein detection method or device and the like. (II) technical scheme In order to achieve the above purpose, the main technical scheme adopted by the invention comprises the following steps: In a first aspect, the invention provides an anti-PR recombinant rabbit monoclonal antibody, which comprises a heavy chain variable region and a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO.4, and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 5. The anti-PR recombinant rabbit monoclonal antibody (PR rabbit source antibody) can be used for immunohistochemical detection, and can identify and detect the expression of PR protein on tumor cells or immune cells with high specificity and high sensitivity. The anti-PR monoclonal antibody is obtained by recombinant expression of mammalian cells. Specifically, the anti-PR recombinant rabbit monoclonal antibody provided by the invention is produced by eukaryotic expression of 293 cells through rabbit hybridoma fusion screening. When the anti-PR monoclonal antibody is prepared, an antigen for immunizing rabbits (New Zealand white rabbits) is a synthetic polypeptide, the amino acid sequence of the synthetic polypeptide is shown as SEQ ID NO. 1, and the synthetic polypeptide is obtained by artificial chemical synthesis. After the rabbit is immunized, a positive hybridoma cell line capable of secreting monoclonal antibodies with high efficiency is obtained through cell fusion and clone screening, a molecular cloning technology is used for obta